Research progress regarding status of human adenovirus infections and serotyping technology / 中华检验医学杂志

There are many serotypes of human adenovirus (HAdV), and different serotypes can cause infections in different systems of the human body, one example being community acquired pneumonia (CAP). However, the clinical symptoms of HAdV infections are similar to those caused by other pathogens. To detect and serotype adenovirus rapidly and accurately is crucial towards the clinical diagnosis, treatment and prevention of the virus. This can facilitates the control of nosocomial infection and epidemiological monitoring. This article briefly reviews the molecular characteristics, epidemiology, and typing of human adenovirus, aiming to help formulate effective treatments towards HAdV infection in a clinical setting.

Suggestions casted to the novel coronavirus nucleic acid amplification test from viral pneumonia pathogenesis / 中华检验医学杂志

An outbreak of Novel Coronavirus (2019-nCoV), results in Coronavirus disease that began in Wuhan, China, has spread rapidly with cases now confirmed in multiple countries. Nucleic acid amplification test (NAAT), represent by reverse-transcriptase polymerase chain reaction (RT-PCR) plays an important role in disease diagnosis and treatment evaluation. The test results by RT-PCR have attracted much attention recently. As understanding to this novel pathogen is still limited, it would be much help to combine the knowledge about its pathogenesis to judge the test results, in addition to review the quality control in laboratory. This review will focus on understanding the specific RT-PCR performance of the 2019-nCoV, under the background of viral pneumonia. The purpose of this review is to add value to NAAT of 2019-nCoV, with combined knowledge of epidemiology, pathogenesis, clinical characteristics and pre-analysis quality control from viral pneumonia.

Knowledges casted to the 2019 novel coronavirus nucleic acid amplification test from pathogenesis of viral pneumonia / 中华检验医学杂志

An outbreak of 2019 novel coronavirus(2019-nCoV), results in novel coronavirus disease that began in Wuhan, China, has spread rapidly with cases now confirmed in multiple countries. Nucleic acid amplification test (NAAT), represent by reverse-transcriptase polymerase chain reaction (RT-PCR) plays an important role in disease diagnosis and treatment evaluation. The test results by RT-PCR have attracted much attention recently. It would be much help to combine the knowledge about its pathogenesis to judge the test results. This review will focus on understanding the specific RT-PCR performance of the 2019-nCoV, with combined knowledge of epidemiology, pathogenesis, clinical characteristics and pre-analysis quality control from viral pneumonia.

Calcium-sensing receptor-mediated L-tryptophan-induced secretion of cholecystokinin and glucose-dependent insulinotropic peptide in swine duodenum

This study aimed to elucidate the effect of tryptophan (Trp) on gut hormone secretion as well as the roles of the calcium-sensing receptor (CaSR) and its downstream signaling pathway in gut hormone secretion by assessing swine duodenal perfusion in vitro. Swine duodenum was perfused with Krebs-Henseleit buffer as a basal solution. Various concentrations (0, 10, and 20 mM) of Trp were applied to investigate its effect on gut hormone secretion. A CaSR antagonist was used to detect the involvement of CaSR and its signal molecules. The 20 mM Trp concentration promoted the secretion of cholecystokinin (CCK) and glucose-dependent insulinotropic peptide (GIP), elevated the mRNA level of CaSR, and upregulated the protein levels of CaSR, protein kinase C (PKC), and inositol trisphosphate receptor (IP3R). However, NPS 2143, an inhibitor of CaSR, attenuated the CCK and GIP release, reduced the mRNA level of CaSR, and decreased the protein levels of CaSR, PKC, and IP3R with 20 mM Trp perfusion. The results indicate that CCK and GIP secretion can be induced by Trp in swine duodenum in vitro, and the effect is mediated by CaSR and its downstream signal molecules PKC and IP3R.

The application of sigma metrics quality models in quality management of clinical biochemistry laboratory / 中华检验医学杂志

Objective To improve the quality of clinical biochemistry laboratory by quality indicators of pre-analytical,analytical,post-analytical phase and the whole process.Methods Analytical Phase:The Sigma values of items were calculated,applying the equation Sigma =(TEa％-Bias％)/CV％.Total allowable error (TEa) is from analyticalal specification defined in WS/T403-2012 of China,Bias％ is from the evaluation results of National Center for Clinical Laboratory (NCCL) trueness verification PT series and CV％ is from internal quality control data during the last 6 months in our lab.Normalized Sigma metrics plot was made to evaluate the analysis performance and the quality control strategies were designed accordingly.The quality goal indexes (QGI) were also calculated to propose improvement measures for items below 6 Sigma.Quality indicators of pre-,post-analytical and whole analytical phase,such as quality of specimen,critical value notification,critical value notification in time,TAT of hs-cTnT,TAT of emergency biochemical items,rewrite of laboratory reports and unacceptable performance in EQA-PT were measured in Sigma metrics too.The Sigma metrics changes before and after taking improvement measures were compared to conform the effectiveness.Results The average Sigma value of 17 biochemical tests was 5.29,of which 8 items (UA,K,ALP,CK,AMY,AST,TG,Na) achieved excellent to world class level (≥ 5 Sigma),6 items (LDH,Cre,TC,ALT,Mg,Glu) achieved marginal to good level (5 ＞ Sigma ≥ 3),BUN performed poorly (3 ＞ Sigma ≥ 2),Ca,TP performed unacceptably (Sigma ＜ 2) with serious quality defects.The Sigma values of unacceptable specimen,critical value notification,critical value notification in time,unacceptable turn around time (TAT) of hs-cTnT,unacceptable turn around time (TAT) of emergency biochemical items,rewrite of laboratory reports,unacceptable performance in EQA-PT were 4.17,3.60,2.75,1.72,3.27,4.52,3.33 respectively,rising to 4.30,4.30,2.90,2.45,3.75,4.80,3.60 accordingly after improvement.Conclusions Sigma metrics is potentially an ideal approach for clinical biochemistry laboratories management,which is helpful to find out problems,put forward improvement measures,and confirm the effectiveness,so as to achieve the purpose of continuous quality improvement.

Detection and clinical analysis of EV71,CA16 and respiratory virus con-infection in patients with hand,foot and mouse disease / 中华检验医学杂志

Objective To explore the situation of respiratory virus co-infection with EV71 and CA16 in patients with hand,foot and mouse disease(HFMD) ,and analyze the influence of co-infection on clinical aspects.Methods From June to October of 2010,there were 348 patients enrolled in the study,with 248 hospitalization cases and 100 mild outpatients.All the patients were diagnosed as HFMD in Beijing You-an Hospital.The viral RNA from the pharynx swab samples were extracted and reversely transcribed by RT-PCR.All the samples were detected with the EV71 and CA16 by real-time fluorescence quantitative PCR.Twelve kinds of respiratory viruses were detected by a commercial multiplex-PCR method.The PCR products were confirmed by electrophoresis.Chi square test was used in the data analysis.Results Of the 348 HFMD patients,36 subjects were detected as positive for respiratory virus co-infection.In the 248 hospitalization cases,111 cases were positive for EV71 or CA16,with eight cases identified with respiratory virus co-infection(7.2%); the other 137 cases were negative for EV71 and CA16,with eleven cases identified with respiratory virus co-infection(7.4%).There was not significant difference between respiratory virus co-infection and the identification of EV71 /CA16(x2 = 0.059,P ＞ 0.05).In the 100 mild outpatients positive for EV71 or CA16,seventeen cases were identified with respiratory virus co-infection(17%).The rate of respiratory virus co-infection in the mild outpatients was much higher than in the severe hospitalization patients(x2 = 4.830,P＜ 0.05).Among the 111 EV71(+) or CA16(+) inpatients,there were 101 cases diagnosed as severe cases(91.0%); similarly,there were 132 cases diagnosed as severe cases(96.4%) among the 137 EV71(-) CA16(-) cases.There was not difference between the identification of EV71/ CA 16 and illness of HFMD(x2 = 3.099,P ＞ 0.05).The leading respiratory virus being identified were HRV A/B,PIV3 and FLU A in the 348 HFMD patients.Conclusions Co-infection with respiratory virus exists in the HFMD patients. However,the respiratory virus infection has no significant influence to the state of HFMD illness.

The investigation of compartmentalization of human immunodeficiency virus infection and CD4~+T lymphocytes between gastric mucosa and peripheral blood from patients with acquired immune deficiency syndrome / 中华传染病杂志

Objective To investigate the compartmentalization of human immunodeficiency virus (HIV)infection and CD4~+ T lymphocytes between gastric mueosa and peripheral blood from patients with acquired immune deficiency syndrome(AIDS).Methods Thirty-five AIDS patients and 10 HIV seronegative subjects were enrolled in this study.The gastric mucosal biopsies were done by gastroscope and peripheral blood samples were collected.The digoxin labeled HIV-1 double-stranded cDNA probes for the long terminal repeat(LTR)and gag gene of HIV-1 genome were prepared by polymerase chain reaction(PCR).In situ hybridization was used to detect HIV infection in gastric mucosal tissues and peripheral blood mononuclear cells(PBMC)of AIDS patients.CD4~+ T lymphocytes were determined by immunohistochemistry method.The data were analyzed using t test.Results HIV positive rates were(1.67±1.48)% in gastric mucosa and(19.37±9.23)% in PBMC of AIDS patients who had not received highly active antiretroviral therapy(HAART).The HIV positive rates in gastric mucosa were not significantly different between AIDS patients without treatment and those treated with HAART(t=-0.996,t=-0.794,t=-0.461;P＞0.05).HIV positive rate on PBMC film preparation in patients who had received HAART for 1-4 years was (4.25±3.47)%,which was significantly lower than untreated group[(19.37±9.23)%](t=3.000,P＜0.05).The positive rate of CD4~+ T lymphocytes was(12.53±8.14)% in gastric mucosal mononuclear cells(MMC)and(19.00±9.55)% in PBMC of AIDS patient who had not received HAART.Even after received HAART for 1-4 years,the positive rate of CD4 ~+ T lymphocytes in MMC were(37.44±18.00)%,which was still lower than control group[(50.35±3.41)%](t=-4.620,P＜0.01);while the positive rate of CD4~+ T lymphocyte in PBMC was similar with that of control group(t=-2.094,P＞0.05).Conclusion The compartmentalization of HIV infection and CD4~+ T lymphocyte counts exists between gastric mucosa and peripheral blood from AIDS patients.

The analysis for human immunodeficiency virus infection in gastric mucosa from acquired immune deficiency syndrome patients / 中华消化杂志

Objective To investigate the distribution and amount of human immunodeficiency virus (HIV) infection in gastric mucosa from untreated acquired immune deficiency syndrome (AIDS) patients and highly active antiretroviral therapy (HAART) treated patients.Methods Thirty-five AIDS patients (14 untreated patients and 21 patients receiving HAART) and 10 HIV-1 seronegative patients with gastrointestinal symptoms were enrolled and examined by upper endoscopy.The labeled HIV-1 double-stranded cDNA probe was a PCR product corresponding to the LTR and gag gene of the HIV-1 genome.HIV in gastric mucosal tissues from AIDS patients was detected using in situ hybridization (ISH) and compared with that in peripheral blood mononuclear cells (PBMC).Results ① No obvious character was found in gastrointestinal symptoms,endoscopy examination and pathology results of AIDS patients.② The expression of HIV gene was mainly detected in the gastric mucosal mononuclear cell (MMC).Other cells were also observed with HIV expression including mucosal epithelial cells,gland epithelial cells and interstitial cells.③There was no difference in HIV expression between sinus ventriculi and gastric body.④ HIV gene expression from AIDS patients was (1.97±3.25)% in gastric mucosa,no difference in HIV gene expression between two groups (P＞0.05).⑤ HIV gene expression in PBMC smear from AIDS patients was (12.38 ± 9.17)%.HIV expreesion in PBMC from patients who had received HAART for 1-4 years were markedly lower than that from patients who had not received HAART (P＜0.05).Conclusions The gastric mueosa is one of HIV infected sites.The potential effect of HAART on the decrease of HIV infected cells in gastric mucosa was unsatisfactory.

Polymorphism analysis on chemokine SDF-1α transcript being the ligand for HIV-1 coreceptor CXCR4 / 中华检验医学杂志

Objective To determine the polymorphism in CXC chemokine SDF-1α transcript and its effects on HIV infection.Methods Three groups of study subjects lived in Hang Kong were recruited:278 HIV-heahhy donors of Chinese origin.49 HIV+Caucasians and 13 Chinese with high risk behavior to HIV but kept uninfected.Genomic DNA and RNA were extracted from eripheral blood mononucleal cell. The PCR and RT-PCR reaction were set up accordingly.Sequence of the SDF-1α promoter,the open reading frame(ORF)and the 3'untranslate region(3'UTR)were analyzed.Two steps PCR reaction using two reverseprimers with mismatched nucleic acid were employed to screen the frequency of a novel mutation. Results equencing analysis from 100 subjects indicated that non mutation happened in tlle promoter and ORF of SDF-1α.A novel mutation Was detected from 3'UTR of SDF-1α.It is a "GA" insertion in "G" rich region near the stop code of SDF-1α.The mutation Was named as SDF-1-3’GA+and submitted to GenBank (AY874118).The mutation happened in three roups.with allele frequency of 15.1% in the healthy Chinese.of 30.7% in the high risk Chinese group.Conclusions our results confirm that SDF-1 genes arerelatively conserved.None noteworthy mutation is identified in the promoter and ORF regions of SDF-1α However.a novel mutation is identified from the 3'UTR of SDF-1α. It would be worthwhile to etermine effect of the novel mutation on HIV infection.

Effect of retroviral vectors carried sense or antisense TGF?1 on human bladder cancer cells in vitro / 中国癌症杂志

Purpose:To investigate the effect of replication defective retroviral vectors carried sense or antisense TGF?1 fragment on the cell cycle regulation and proliferation of human bladder cancer. Methods:The replication defective retroviral vectors that integrated sense or antisense bioactive fragment of transforming growth factor?1 were constructed,and named as pRevT? and pRevT?-AS respectively. The influence of each vector on the cell proliferation,clone-formation and alteration of cell cycle of bladder cancer cell line EJ were observed in vitro.Results:The titre of pRevT? and pRevT?-AS were 0.84,0.88?10 5 CFU/ml respectively,the vectors integrated to EJ cells and expressed efficiently. Inhibition TGF?1 gene expression reduced proliferation and clone-formation rates of EJ cells. The G 0 /G 1 stage ratios in the antisense TGF?1-transfected EJ cells were increased,simultaneously,the S stage ratios were decreased. Conclusions:The antisense TGF?1 vector can reduce the expression of endogenous TGF?1 in EJ cells,induce G 1 stage arrest and inhibit proliferative growth in vitro.